Detection of drug resistance in Mycobacterium tuberculosis: Methods, principles and applications

Gupta, Anamika; Anupurba, Shampa

doi:10.1016/j.ijtb.2015.02.003

Cited by 13 publications

(10 citation statements)

References 65 publications

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“…En los últimos años se han realizado estudios relacionados con la resistencia a medicamentos que han permitido el desarrollo de métodos fenotípicos y genotípicos 25 .…”

Section: Discussionunclassified

“…La INH actúa inhibiendo la biosíntesis de los ácidos micólicos de la pared celular, e ingresa en el Mycobacterium tuberculosis como un profárma-co por difusión pasiva y es activada por la enzima catalasaperoxidasa codificada por katG, para generar radicales libres, que atacan a continuación múltiples objetivos en las células 8 . Se ha reportado en varios estudios que la resistencia a isoniacida está relacionada con mutaciones específicas en varios genes importantes de Mycobacterium tuberculosis, existiendo variantes genéticas tales como: katG (S315T, S315N, S315D, S315I, S315R, V61G, Q247H, A62T, G383A, C701G, G1388T), inhA (S94A, I21V, I21T, I47T, I194T, 3, 258, 190) y su región promotora (-15 C-T, -8 T-C, T-A ó T-G,-17 G-T), ahpC, nhd, kasA, oxyR, furA, fabG1 [9][10][11][12][13][14][15][16][17][18][19][20][21] , de igual manera, se ha podido mostrar que la aplicación de PCR-RFLP del gen katG permite la identificación eficiente de aislados resistentes a isoniacida pudiendo emplearse como una prueba rápida de detección [22][23][24][25] .…”

Section: Introductionunclassified

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Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Sánchez-Domínguez

Nicola-Salas

Morey-León

2018

Infect

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Cómo citar este artículo: J. Sánchez-Domínguez, et al. Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP. Infectio 2018; 22(4): 178-184 ResumenObjetivo: determinar la mutación S315T del gen katG en aislados de Mycobacterium tuberculosis resistentes a isoniacida mediante la técnica PCR-RFLP. Materiales y métodos: A partir de 68 aislados de Mycobacterium tuberculosis se realizó el análisis de polimorfismo en productos de amplificación de 1054 y 630 pb que contenían la mutación S315T del gen katG mediante PCR-RFLP empleando las enzimas de restricción MspI y SatI. Mediante SPSS se determinó sensibilidad, especificidad, valores predictivo positivo y negativo, coeficientes de probabilidad positivo y negativo. Resultados: El 74,46% de aislados fenotípicamente resistentes y 4,76% fenotípicamente sensibles presentaron la mutación del gen katG S315T. La PCR-RFLP para S315T del gen katG presentó 85,4% de sensibilidad y 95,2% de especificidad con MspI y 85,4% de sensibilidad y 94,4% de especificidad con SatI. Discusión: La PCR-RFLP tiene una alta capacidad resolutiva que depende de la enzima que se emplee como se observó en estudios previos. La presencia de la mutación S315T en pacientes vírgenes al tratamiento sugiere la circulación de aislados resistentes a Isoniacida. Conclusión: La PCR-RFLP resultó una alternativa válida y rápida para el diagnóstico de la resistencia a isoniacida, mediante la detección de la mutación S315T del gen katG en comparación con el método convencional de las proporciones.Palabras Clave: Mutaciones, Polimorfismo, Isoniacida, PCR-RFLP, Mycobacterium tuberculosis, katG. Determination of S315T mutation within the katG gene in isoniazid-resistant Mycobacterium tuberculosis isolate by PCR-RFLP AbstractObjetive: To determine the S315T mutation of the katG gene in Mycobacterium tuberculosis isoniazid-resistant isolates by PCR-RFLP. Materials and Methods: Polymorphism analysis of 1054 and 630 bp products containing the S315T mutation of the katG gene was performed by PCR-RFLP using the MspI and SatI restriction enzymes from 68 Mycobacterium tuberculosis isolates. Sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratio were determined using SPSS. Results: 74.46% of isoniazid-resistant and 4.76% of isoniazid-sensitive isolates showed the S315T mutation in katG gene. The PCR-RFLP for S315T of the katG gene had 85.4% sensitivity and 95.2% specificity with MspI and 85.4% sensitivity and 94.4% specificity with SatI. Discussion: The PCR-RFLP has a high resolutive capacity that depends on the enzyme that is used as it was observed in previous studies. The presence of the S315T mutation in treatment-naive patients suggests the circulation of isolates resistant to isoniazid. Conclusion: PCR-RFLP is a valid and rapid alternative for the diagnosis of isoniazid resistance, by detection of S315T mutation in the katG gene compared to the conventional method of proportions.

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“…En los últimos años se han realizado estudios relacionados con la resistencia a medicamentos que han permitido el desarrollo de métodos fenotípicos y genotípicos 25 .…”

Section: Discussionunclassified

Section: Introductionunclassified

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Sánchez-Domínguez

Nicola-Salas

Morey-León

2018

Infect

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“…ATP-binding transport protein family (ABC transporters) [27,33] Isoniazid Rv1634 Major facilitator superfamily (MFS), [33] Pyrazinamide Rv1667c ATP-binding transport protein family (ABC transporters) [32] Isoniazid Rv1686c-Rv1687c ATP-binding transport protein family (ABC transporters) [33] Cycloserine Rv1704c (cycA) Amino acid permease family (APC family) [41] Streptomycin Rv1877 Major facilitator superfamily (MFS) [24] O oxacin Rv2209 Probable conserved integral membrane protein [20] Isoniazid, Spectinomycin Tetracycline Rv2333c (stp) Major facilitator superfamily (MFS) [42] Isoniazid, Ethambutol Rv2459 (jefA) Major facilitator superfamily (MFS) [20,37,43] O oxacin Rv2477c ATP-binding transport protein family (ABC transporters) [20] Streptomycin, Cipro oxacin Rv2686c, Rv2687c, Rv2688c…”

Section: Rv1258cmentioning

confidence: 99%

Efflux pump genes variants in MDR TB strains with discrepant phenotype-genotype correlations may further guide drug resistance interpretation

Hasan¹,

Razzak²,

Kanji³

et al. 2023

Preprint

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Introduction: Whole genome analysis of Mycobacterium tuberculosis (MTB) is an increasingly important method of identification of multi-drug resistance (MDR) in clinical isolates. It is based on the identification of single nucleotide variants (SNVs) in genes associated with resistance. However, there remain gaps in our understanding of phenotype – genotype correlation between strains. Efflux pumps contribute to drug resistance and here we studied SNVs in key efflux pump genes (EP) to investigate their association with resistance. Methods: Whole genome data of 2221 MTB isolates comprising 1432 susceptible and 789 drug resistant strains were downloaded from ReSeqTB database. MTB lineage and resistance genotyping analysis was performed using an in-house bioinformatics pipeline, MTB-VCF. SNVs in 47 EP genes were categorized according to their SIFT/Polyphen scores. Results: We identified variants unique to EP in DR isolates. SIFT/Polyphen effect analysis determined 38 high impact SNVs across twenty EP genes (EP) to be present in these 789 genomes. SNVs were not associated with MTB lineages. The EPs with SNVs in DR isolates were Rv1819, Rv0194, Rv0507, Rv2333c, Rv3728, Rv3823, Rv1250, Rv1273, Rv1458, Rv1634, Rv1217, Rv1218, Rv0450, Rv0676c, Rv0191, Rv3008, Rv3756, Rv2688, Rv1704 and Rv1877. Examination of 52 isolates with discrepant phenotype-genotypes resistance comprising, MDR, pre-XDR and XDR strains revealed SNVs in EP associated with RIF and INH (Rv0194, Rv1217_1218, Rv1819, Rv0450, Rv1458, Rv0507), and those associated with fluoroquinolone (Rv1634 and Rv2688) resistance. Conclusions: We identified SNVs in efflux pumps which could contribute to resistance in MTB strains. It may be important to consider these as part of MTB genome-based resistance interpretation. Functional studies combined with GWAS and RNA profiling would further confirm these findings.

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“…Quick diagnosis and cure are crucial as when TB patients are not diagnosed and cured quickly, they may unknowingly spread their infection to their families and communities and exacerbating the epidemic [6]. The century-old smear microscopy and culture methods are exasperatingly insensitive (sensitivity between 50-52%) and time-consuming, respectively [7,8].…”

Section: Scenario In Tb Diagnosticsmentioning

confidence: 99%

Imminent Weapons in the Field of Tuberculosis Diagnostics and Management

Nema¹

2017

IMPH

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Tuberculosis (TB) has always been a challenge for clinicians and researchers. A lot have been done for its management and lot more is awaited. Diagnosing TB became faster with the advent and validation of Xpert ® MTB/RIF (GeneXpert MTB/RIF or Xpert) and improvements over these instruments are coming to help to resolve bottlenecks. Similarly, better drugs with shorter treatment durations are long awaited and there has been some progress on this front too. This short review discusses recent advances from these fields.

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Detection of drug resistance in Mycobacterium tuberculosis: Methods, principles and applications

Cited by 13 publications

References 65 publications

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Efflux pump genes variants in MDR TB strains with discrepant phenotype-genotype correlations may further guide drug resistance interpretation

Imminent Weapons in the Field of Tuberculosis Diagnostics and Management

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